RNA amplification chip with parallel microchannels and droplet positioning using capillary valves

Abstract

We present results from the MicroActive project which develops an instrument for molecular diagnostics. The instrument is first tested for patient screening for a group of viruses causing cervical cancer. Two disposable polymer chips with reagents stored on-chip are developed and will be inserted into the instrument for each patient sample analysis. The first chip will perform nucleic acid extraction from patient epithelial cervical cells, while mRNA amplification and fluorescent detection takes place in the second chip. This paper reports results on the amplification chip. Purified sample is inserted into the chip and split into ten smaller droplets for simultaneous amplification and detection of ten viruses. The droplets move in parallel channels, each with two chamber extensions containing dried reagents. Experimental results on parallel droplet movement using one external pump combined with hydrophobic restrictions show that the parallel droplet positions can be controlled. There are four valves with increasing burst pressures between 800 and 4,500 Pa in each parallel channel, positioning the droplets in metering zones and reaction chambers. The re-hydration times for the dried reagents in micro chambers have been monitored. After sample insertion, uniform concentration of the reagents in the droplet was reached after respectively 60 s and 10 min. These times are acceptable for successful amplification. Finally we show positive amplification of HPV type 16 viruses in a micro chamber.